Consulta: subjectFacets:"QPCR"
Registros recuperados: 34
Data/hora: 01/06/2024 08:33:49

Chilean IPNV isolates: Robustness analysis of PCR detection
Provedor de dados: 69
Autores: Jorquera,Esteban; Morales,Paz; Tapia,David; Torres,Pamela; Eissler,Yoanna; Espinoza,Juan C; Conejeros,Pablo; Kuznar,Juan.
Background: The genomes of several infectious pancreatic necrosis viruses (IPNVs) isolated in Chile were sequenced with a single amplification approach for both segments A and B. The resulting sequences were then used to determine the conservation of the primer-binding regions used in polymerase chain reaction (PCR)-based diagnostic methods proposed in the literature. Thus, the robustness of each technique was studied, particularly the eventual effect of further mutations within the primer-binding sites. Results: On analysis, most methods currently used to detect Chilean IPNV varieties were deemed adequate. However, the primers were designed to be genogroup specific, implying that most detection methods pose some risk of detecting all strains prevalent in...
Tipo: Journal article
Palavras-chave: IPNV detection; Mismatch's Tm analysis; QPCR.
Ano: 2016
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000200005

Expression analysis in response to drought stress in soybean: shedding light on the regulation of metabolic pathway genes
Provedor de dados: 74
Autores: Guimarães-Dias,Fábia; Neves-Borges,Anna Cristina; Viana,Antonio Americo Barbosa; Mesquita,Rosilene Oliveira; Romano,Eduardo; Grossi-de-Sá,Maria de Fátima; Nepomuceno,Alexandre Lima; Loureiro,Marcelo Ehlers; Alves-Ferreira,Márcio.
Metabolomics analysis of wild type Arabidopsis thaliana plants, under control and drought stress conditions revealed several metabolic pathways that are induced under water deficit. The metabolic response to drought stress is also associated with ABA dependent and independent pathways, allowing a better understanding of the molecular mechanisms in this model plant. Through combining an in silico approach and gene expression analysis by quantitative real-time PCR, the present work aims at identifying genes of soybean metabolic pathways potentially associated with water deficit. Digital expression patterns of Arabidopsis genes, which were selected based on the basis of literature reports, were evaluated under drought stress condition by Genevestigator. Genes...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Glycine max; Drought resistance; QPCR; Metabolic pathway; Bioinformatics.
Ano: 2012
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572012000200004

Ciguatoxin Occurrence in Food-Web Components of a Cuban Coral Reef Ecosystem: Risk-Assessment Implications
Provedor de dados: 5
Autores: Díaz-asencio, Lisbet; Clausing, Rachel J.; Vandersea, Mark; Chamero-lago, Donaida; Gómez-batista, Miguel; Hernández-albernas, Joan I.; Chomérat, Nicolas; Rojas-abrahantes, Gabriel; Litaker, Wayne; Tester, Patricia; Diogène, Jorge; Alonso-hernández, Carlos M.; Dechraoui Bottein, Marie-yasmine.
In Cuba, ciguatera poisoning associated with fish consumption is the most commonly occurring non-bacterial seafood-borne illness. Risk management through fish market regulation has existed in Cuba for decades and consists of bans on selected species above a certain weight; however, the actual occurrence of ciguatoxins (CTXs) in seafood has never been verified. From this food safety risk management perspective, a study site locally known to be at risk for ciguatera was selected. Analysis of the epiphytic dinoflagellate community identified the microalga Gambierdiscus. Gambierdiscus species included six of the seven species known to be present in Cuba (G. caribaeus, G. belizeanus, G. carpenteri, G. carolinianus, G. silvae, and F. ruetzleri). CTX-like...
Tipo: Text
Palavras-chave: Caribbean; Ciguatoxicity; QPCR; Trophic transfer; Ish; Food safety; Food security; Science-based management; Foodborne disease.
Ano: 2019
URL: https://archimer.ifremer.fr/doc/00597/70947/69189.pdf

Environmental DNA for detecting Bulinus truncatus : A new environmental surveillance tool for schistosomiasis emergence risk assessment
Provedor de dados: 5
Autores: Mulero, Stephen; Boissier, Jérôme; Allienne, Jean‐françois; Quilichini, Yann; Foata, Joséphine; Pointier, Jean‐pierre; Rey, Olivier.
Under ongoing climate changes, the development of large‐scale monitoring tools for assessing the risk of disease emergence constitutes an urging challenge. This is particularly the case for snail‐borne diseases such as the urogenital bilharziasis that emerged in Corsica and threat European countries. The expansion of this tropical disease mainly relies on the local presence of competent snail hosts such as Bulinus truncatus. Unfortunately, very little is known about the actual repartition of freshwater snails worldwide which makes new emergences difficult to predict. In this study, we developed two ready‐to‐use environmental DNA‐based methods for assessing the distribution of B. truncatus from water samples collected in the field. We used two approaches, a...
Tipo: Text
Palavras-chave: Bulinus truncatus; Corsica; DdPCR; Environmental DNA; Environmental monitoring; QPCR; Schistosomiasis.
Ano: 2020
URL: https://archimer.ifremer.fr/doc/00593/70554/68723.pdf

Assessment of Ciguatera and Other Phycotoxin-Related Risks in Anaho Bay (Nuku Hiva Island, French Polynesia): Molecular, Toxicological, and Chemical Analyses of Passive Samplers
Provedor de dados: 5
Autores: Roué, Mélanie; Smith, Kirsty F.; Sibat, Manoella; Viallon, Jérôme; Henry, Kévin; Ung, André; Biessy, Laura; Hess, Philipp; Darius, Hélène Taiana; Chinain, Mireille.
Ciguatera poisoning is a foodborne illness caused by the consumption of seafood contaminated with ciguatoxins (CTXs) produced by dinoflagellates from the genera Gambierdiscus and Fukuyoa. The suitability of Solid Phase Adsorption Toxin Tracking (SPATT) technology for the monitoring of dissolved CTXs in the marine environment has recently been demonstrated. To refine the use of this passive monitoring tool in ciguateric areas, the effects of deployment time and sampler format on the adsorption of CTXs by HP20 resin were assessed in Anaho Bay (Nuku Hiva Island, French Polynesia), a well-known ciguatera hotspot. Toxicity data assessed by means of the mouse neuroblastoma cell-based assay (CBA-N2a) showed that a 24 h deployment of 2.5 g of resin allowed...
Tipo: Text
Palavras-chave: Ciguatera monitoring; Gambierdiscus; Ciguatoxins; SPATT passive samplers; HP20 resin; CBA-N2a; LC-MS; MS; WS artificial substrate; QPCR; HTS metabarcoding.
Ano: 2020
URL: https://archimer.ifremer.fr/doc/00629/74143/73670.pdf

Development of a qPCR Method for the Identification and Quantification of Two Closely Related Tuna Species, Bigeye Tuna (     Thunnus obesus     ) and Yellowfin Tuna (     Thunnus albacares     ), in Canned Tuna
Provedor de dados: 5
Autores: Bojolly, Daline; Doyen, Perine; Le Fur, Bruno; Christaki, Urania; Verrez-bagnis, Veronique; Grard, Thierry.
Bigeye tuna (Thunnus obesus) and yellowfin tuna (Thunnus albacares) are among the most widely used tuna species for canning purposes. Not only substitution but also mixing of tuna species is prohibited by the European regulation for canned tuna products. However, as juveniles of bigeye and yellowfin tunas are very difficult to distinguish, unintentional substitutions may occur during the canning process. In this study, two mitochondrial markers from NADH dehydrogenase subunit 2 and cytochrome c oxidase subunit II genes were used to identify bigeye tuna and yellowfin tuna, respectively, utilizing TaqMan qPCR methodology. Two different qPCR-based methods were developed to quantify the percentage of flesh of each species used for can processing. The first one...
Tipo: Text
Palavras-chave: Tuna; Authentication; Quantification; Canned products; TaqMan; QPCR; Species identification; Bigeye tuna (Thunnus obesus); Yellowfin tuna (Thunnus albacares).
Ano: 2017
URL: https://archimer.ifremer.fr/doc/00371/48217/48403.pdf

Differential Mortality and High Viral Load in Naive Pacific Oyster Families Exposed to OsHV-1 Suggests Tolerance Rather than Resistance to Infection
Provedor de dados: 5
Autores: Agnew, M. Victoria; Friedman, Carolyn S.; Langdon, Christopher; Divilov, Konstantin; Schoolfield, Blaine; Morga, Benjamin; Degremont, Lionel; Dhar, Arun K.; Kirkland, Peter; Dumbauld, Brett; Burge, Colleen A.
Pacific oysters, Crassostrea gigas, are one of the most productive aquaculture species in the world. However, they are threatened by the spread of Ostreid herpesvirus-1 (OsHV-1) and its microvariants (collectively “µvars”), which cause mass mortalities in all life stages of Pacific oysters globally. Breeding programs have been successful in reducing mortality due to OsHV-1 variants following viral outbreaks; however, an OsHV-1-resistant oyster line does not yet exist in the United States (US), and it is unknown how OsHV-1 µvars will affect US oyster populations compared to the current variant, which is similar to the OsHV-1 reference, found in Tomales Bay, CA. The goals of this study were to investigate the resistance of C. gigas juveniles produced by the...
Tipo: Text
Palavras-chave: OsHV-1; Mu vars; Oyster; Virus; Tolerance; Virulence; QPCR.
Ano: 2020
URL: https://archimer.ifremer.fr/doc/00665/77739/79862.pdf

Rearing practices identified as risk factors for ostreid herpesvirus 1 (OsHV-1) infection in Pacific oyster Crassostrea gigas spat
Provedor de dados: 5
Autores: Normand, Julien; Blin, Jean-louis; Jouaux, Aude.
Early detection of Pacific oyster spat infected with ostreid herpesvirus 1 (OsHV-1) could prevent introduction of OsHV-1-infected individuals into farming areas or onshore rearing facilities, thus reducing the risk of infection of naïve oysters in such production systems. Experiments were conducted on several hundred oyster spat provided by producers in order to examine whether early rearing practices could be considered as potential risk factors for (1) OsHV-1 infection as detected by molecular methods and (2) spat mortality experimentally induced through thermal challenge. Spat groups collected on oyster beds and hatchery spat reared in growout areas during summer exhibited higher viral DNA contamination and mortalities during the trial than spat kept in...
Tipo: Text
Palavras-chave: Oyster spat; Epidemiology; OsHV-1; QPCR; Thermal challenge.
Ano: 2014
URL: https://archimer.ifremer.fr/doc/00203/31407/29797.pdf

Immune priming and portal of entry effectors improve response to vibrio infection in a resistant population of the European abalone
Provedor de dados: 5
Autores: Dubief, Bruno; Nunes, Flavia; Basuyaux, Olivier; Paillard, Christine.
Since 1997, populations of the European abalone Haliotis tuberculata suffer mass mortalities attributed to the bacterium Vibrio harveyi. These mortalities occur at the spawning season, when the abalone immune system is depressed, and when temperatures exceed 17 °C, leading to favorable conditions for V. harveyi proliferation. In order to identify mechanisms of disease resistance, experimental successive infections were carried out on two geographically distinct populations: one that has suffered recurrent mortalities (Saint-Malo) and one that has not been impacted by the disease (Molène). Furthermore, abalone surviving these two successive bacterial challenges and uninfected abalone were used for several post-infection analyses. The Saint-Malo population...
Tipo: Text
Palavras-chave: Immunity; Hemocyte; Abalone; Disease; Extracellular products; Immune priming; Vibrio harveyi; Flow cytometry; Resistance; Phagocytosis; Bacterial growth; QPCR; Gill.
Ano: 2017
URL: http://archimer.ifremer.fr/doc/00357/46800/46667.pdf

An updated assessment of     Symbiodinium     spp. that associate with common scleractinian corals from Moorea (French Polynesia) reveals high diversity among background symbionts and a novel finding of clade B
Provedor de dados: 5
Autores: Rouze, Heloise; Lecellier, Gael J.; Saulnier, Denis; Planes, Serge; Gueguen, Yannick; Wirshing, Herman H.; Berteaux-lecellier, Veronique.
The adaptative bleaching hypothesis (ABH) states that, depending on the symbiotic flexibility of coral hosts (i.e., the ability of corals to “switch” or “shuffle” their algal symbionts), coral bleaching can lead to a change in the composition of their associated Symbiodinium community and, thus, contribute to the coral’s overall survival. In order to determine the flexibility of corals, molecular tools are required to provide accurate species delineations and to detect low levels of coral-associated Symbiodinium. Here, we used highly sensitive quantitative (real-time) PCR (qPCR) technology to analyse five common coral species from Moorea (French Polynesia), previously screened using only traditional molecular methods, to assess the presence of...
Tipo: Text
Palavras-chave: Corals; French polynesia; Clade B; Symbiodinium; QPCR; Flexibility; Generalist; Faithful clade.
Ano: 2017
URL: http://archimer.ifremer.fr/doc/00365/47652/47683.pdf

First comparison of French and Australian OsHV-1 mu vars by bath exposure
Provedor de dados: 5
Autores: Burge, Colleen A.; Reecez, Kimberly S.; Dhar, Arun K.; Kirkland, Peter; Morga, Benjamin; Degremont, Lionel; Faury, Nicole; Wippel, Bryanda J. T.; Macintyre, Alanna; Friedman, Carolyn S.
Economically devastating mortality events of farmed and wild shellfish due to infectious disease have been reported globally. Currently, one of the most significant disease threats to Pacific oyster Crassostrea gigas culture is the ostreid herpesvirus 1 (OsHV-1), in particular the emerging OsHV-1 microvariant genotypes. OsHV-1 microvariants (OsHV-1 mu vars) are spreading globally, and concern is high among growers in areas unaffected by OsHV-1. No study to date has compared the relative virulence among variants. We provide the first challenge study comparing survival of naive juvenile Pacific oysters exposed to OsHV-1 mu vars from Australia (AUS mu var) and France (FRA mu var). Oysters challenged with OsHV-1 mu vars had low survival (2.5%a exposed to AUS...
Tipo: Text
Palavras-chave: Ostreid herpesvirus 1; Microvariant; OsHV-1 mu vars; POMS; Pacific oyster; Crassostrea gigas; Viral disease; QPCR; Alkalinity; Emerging infectious diseases.
Ano: 2020
URL: https://archimer.ifremer.fr/doc/00631/74277/73889.pdf

Signature d'expression de gènes, Marqueur prédictif de capacités de survie chez l'huître Crassostrea gigas
Provedor de dados: 5
Autores: Bachere, Evelyne; Degremont, Lionel; Piquemal, David; Roman, Bruno; Noguier, Florian.
Dans le contexte des mortalités massives d'huîtres Crassostrea gigas, les mécanismes moléculaires et les bases génétiques impliquées dans la capacité de survie des huîtres ont été explorées par des approches de génomique à très haut débit. Ces travaux se sont centrés dans un premier temps sur la survie à des infections par des Vibrio virulents pour lesquels des infections expérimentales étaient reproductibles et standardisées en laboratoire. Ces analyses ont conduit à la mise en évidence d'une série de gènes dont l’expression peut prédire la capacité des huîtres à survivre aux infections à vibrios (Rosa et al. 2012). Depuis 2013, les travaux ont permis la validation de la signature de survie qui a pu être corrélée avec les capacités de survie des huîtres...
Tipo: Text
Palavras-chave: Huître creuse; Signature d'expression de gènes; QPCR; Biostatistique; Capacité de survie; Phénotypage; Héritabilité; Pacific oyster; Gene expression signature; QPCR; Biostatistics; Survival capacity; Phenotyping; Heritability.
Ano: 2017
URL: http://archimer.ifremer.fr/doc/00434/54551/55925.pdf

Detection of Salmonella spp. by Conventional Bacteriology and by Quantitative Polymerase-Chain Reaction in Commercial Egg Structures
Provedor de dados: 89
Autores: Moraes,DMC; Duarte,SC; Bastos,TSA; Rezende,CLG; Leandro,NSM; Café,MB; Stringhini,JH; Andrade,MA.
Abstract Conventional bacteriology techniques and quantitative polymerase-chain reaction (qPCR) were applied to the eggshell, albumen, and yolk of washed and unwashed commercial white and brown eggs, to detect Salmonella spp. Pooled samples of eggshells, albumen, and yolk of white and brown eggs were collected at the poultry house and at the egg-storage room. Salmonella spp. was detected by conventional bacteriology in 5.4% (21/387) of analyzed samples and in 16% (68/387) by qPCR. In the 114 unwashed white eggs samples of eggshell, albumen and yolk, the bacterium was identified in 2.6% of the eggs (3/114) by conventional bacteriology and in 13.2% (15/114) by qPCR. In the 90 samples of washed eggs, 6.7% (6/90) were contaminated as detected by conventional...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: QPCR; Eggshells; Albumen; Yolk.
Ano: 2016
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-635X2016000100117

Effects of in Ovo Infusion of Probiotic Strains on Performance Parameters, Jejunal Bacterial Population and Mucin Gene Expression in Broiler Chicken
Provedor de dados: 89
Autores: Majidi-Mosleh,A; Sadeghi,AA; Mousavi,SN; Chamani,M; Zarei,A.
ABSTRACT The objective of the present study was to evaluate the effects of in ovo infusion of probiotic strains (Bacillus subtilis, Enterococcus faecium, and Pediococcus acidilactici ) on jejunal microbial population and mucin gene expression in broiler chicken. In a completely randomized design, 0.5 ml of mediums containing 107 cfu of different probiotic strains, was administered into amniotic fluid of the 480 Cobb fetus (day 18 of incubation), with four treatments, five replicates with twenty four eggs each. For mucin gene expression, samples from the jejunum were taken on day 21 of incubation and day 3 post-hatch. Microbial profile was determined for total lactobacillus and E. coli by sampling jejunal contents on days 1 and 3 of age. Expression of the...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: In ovo infusion; Jejunum; Microbial population; QPCR; Probiotic.
Ano: 2017
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-635X2017000500097

Relative expression of CYP and Arabinogalactan genes in soybean seed coats
Provedor de dados: 124
Autores: Bahry,Carlos André; Venske,Eduardo; Perboni,Anelise Tessari; Zimmer,Paulo Dejalma.
ABSTRACT  The aim of this study was to evaluate the relative expression of two candidate genes, CYP (cyclophilin) and Arabinogalactan, possibly involved with seed quality, in contrasting soybean seed coats. The following genotypes were used: BMX Potência RR and CD 202, both with yellow coats, and TP and IAC, with black coats. The relative expression of the genes was evaluated by qPCR in seven development phases of the coats of the four genotypes at 25; 30; 35; 40; 45; 50 and 55 days after anthesis. The design was completely randomized, with three replicates. The data were submitted to analysis of variance and the means were compared via the Scott Knott test, at 5% probability. The CYP gene showed greater expression in the intermediate stages of development...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Contrasting genotypes; Candidate genes; Cyclophilin; QPCR.
Ano: 2018
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1806-66902018000100078

Cuantificación de maíz genéticamente modificado mediante las técnicas de qPCR y dPCR
Provedor de dados: 115
Autores: Gutiérrez-Angoa,Lizbeth E.; Castillo-Durán,Luis C.; Gómez-Castelo,Blanca E.; Acatzi-Silva,Abraham I.
A partir de la emisión de los permisos de liberación al ambiente de maíz (Zea mays L.) genéticamente modificado (GM) en el año 2009, en México fue necesario realizar la detección, identificación y cuantificación de organismos genéticamente modificados (OGM) de los cultivos en el país. El objetivo del presente estudio fue validar la técnica de cuantificación absoluta de mezclas de hoja en fracción masa de maíz GM a través de la técnica de la reacción en cadena de la polimerasa (PCR) en tiempo real (qPCR) y digital (dPCR). Mezclas de hojas de maíz modificado con el evento MON810 se prepararon con hojas de maíz convencional, fueron analizadas con la técnica de qPCR, y como calibrantes se usaron un plásmido de referencia certificado y ADN obtenido de una hoja...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: QPCR; DPCR; Zea mays L.; OGM.
Ano: 2015
URL: http://www.scielo.org.mx/scielo.php?script=sci_arttext&pid=S1405-31952015000400003

Detection and quantification of human adenovirus genomes in Acanthamoeba isolated from swimming pools
Provedor de dados: 42
Autores: STAGGEMEIER,RODRIGO; ARANTES,THALITA; CAUMO,KARIN S.; ROTT,MARILISE B.; SPILKI,FERNANDO R.
ABSTRACT Acanthamoeba is the most common free-living environmental amoeba, it may serve as an important vehicle for various microorganisms living in the same environment, such as viruses, being pathogenic to humans. This study aimed to detect and quantify human adenoviruses (HAdV) in Acanthamoebas isolated from water samples collected from swimming pools in the city of Porto Alegre, Southern Brazil. Free-living amoebae of the genus Acanthamoeba were isolated from water samples, and isolates (n=16) were used to investigate the occurrence of HAdVs. HAdV detection was performed by quantitative real-time polymerase chain reaction (qPCR). HAdVs were detected in 62.5% (10/16) of Acanthamoeba isolates, ranging from 3.24x103 to 5.14x105 DNA copies per milliliter...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Acanthamoeba; Free-living amoebae; Human adenovirus; QPCR; Swimming pools.
Ano: 2016
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652016000200635

Comparative validation using quantitative real-time PCR (qPCR) and conventional PCR of bovine semen centrifuged in continuous density gradient
Provedor de dados: 45
Autores: Resende,M.V.; Lucio,A.C.; Perini,A.P.; Oliveira,L.Z.; Almeida,A.O.; Alves,B.C.A.; Moreira-Filho,C.A.; Santos,I.W.; Hossepian de Lima,V.F.M.
The objective of the present study was to determine the sperm enrichment with X-bearing spermatozoa, after one centrifugation in a Percoll or OptiPrep continuous density gradient, using quantitative real-time polymerase chain reaction (qPCR) of sperm DNA and resultant in vitro-produced bovine embryos by PCR. Frozen/thawed sperm was layered on density gradients and the tubes were centrifuged. Supernatants were gently aspirated and the sperm recovered from the bottom of the tubes. Cleavage and blastocyst rates were determined through in vitro production of embryos and PCR was performed to identify the embryos' genetic sex. A difference in blastocyst rate was found in the Percoll treatment compared to OptiPrep (P&lt;0.05). The percentage of female embryos in...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Bovine; Sperm sexing; Centrifugation; Embryo sexing; QPCR; PCR.
Ano: 2011
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352011000300002

Occurrence of Mycobacterium avium subsp. paratuberculosis in coalho cheese in the State of Pernambuco, Brazil
Provedor de dados: 45
Autores: Albuquerque,P.P.F.; Cezar,R.D.S.; Pinheiro Junior,J.W.; Grazielle Nascimento,G.; Santos,A.S.; Mota,R.A.
ABSTRACT Paratuberculosis is a chronic and incurable disease that affects ruminants and other domestic animals. It is caused by Mycobacterium avium subsp. paratuberculosis (MAP) that may also be involved in some human diseases such as Crohn's disease, type 1 diabetes, sarcoidosis, multiple sclerosis, and Hashimoto's thyroiditis. The objective of this study was to investigate the occurrence of MAP DNA in samples of artisanal coalho cheese purchased in the State of Pernambuco. Forty samples of coalho cheese submitted to the Real Time Polymerase Chain Reaction (qPCR) technique were analyzed for the detection of the MAP region IS900. 11 (27.5%) were positive with a mean of 195.9 MAP colony forming unit (CFU) per gram of each sample, with a minimum of 30.3...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Bovine; IS900; QPCR; Paratuberculosis.
Ano: 2019
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352019000601917

Molecular investigation of Neorickettsia risticii in trematodes and snails in a region with serological evidence of this agent in horses, state of Rio de Janeiro
Provedor de dados: 45
Autores: Costa,R.L.; Vitari,G.L.V.; Silva,C.B.; Peckle,M.P.; Pires,M.S.; Brandolini,S.V.P.B.; Pinheiro,J.; Massard,C.L.; Santos,H.A.
ABSTRACT In Brazil, some studies have indicated that Neorickettsia risticii circulates in horses, but it is unclear which are the possible intermediate vectors of this bacterium in the country. The aim of this study was to use molecular techniques in order to analyze the presence of N. risticii in snails and larval stages of trematodes in farms in a region with a history of seroreactive horses towards this bacterium, in Rio de Janeiro, Brazil. Convenience sampling was used in the studied region. The collected snails were exposed to incandescent light (60W) for 2-4 hours in order to investigate trematodes in larval forms. Deoxyribonucleic acid (DNA) was extracted from snail tissue and trematode. Real-time PCR (qPCR) technique was used to investigate the...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Equine neorickettsiosis; Molecular diagnosis; QPCR; Intermediate hosts.
Ano: 2016
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352016000601470

Accuracy of quantitative polymerase chain reaction in samples of frozen and paraffin-embedded healthy skin for the diagnosis of canine visceral leishmaniasis
Provedor de dados: 45
Autores: Campos,M.P.; Madeira,M.F.; Silva,D.A.; Solcà,M.S.; Espíndola,O.M.; Mendes Júnior,A.A.V.; Figueiredo,F.B.
ABSTRACT The purpose of the present work was to evaluate the accuracy of quantitative polymerase chain reaction (qPCR) performed on samples of fresh frozen tissue (FT) and formalin-fixed, paraffin-embedded (FFPE) healthy skin. This is a validation study conducted with samples from 46 dogs from an endemic area in Brazil. After sample collection, DNA extractions were conducted using commercial kits and qPCR was oriented to kinetoplast DNA (kDNA) targets of the Leishmania infantum species. The results obtained for the FFPE samples showed 63.6% sensitivity and 77.1% specificity, whereas those obtained for the FT samples showed 100% and 48.6%, respectively. Poor agreement was observed for the results of the qPCR technique with FT and FFPE samples. Our results...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Diagnosis; QPCR; Canine visceral leishmaniasis.
Ano: 2017
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352017000601443

PCR em tempo real para detecção do vírus da doença de Aujeszky
Provedor de dados: 45
Autores: Fonseca Júnior,A.A.; Cotorello,A.C.; Dias,N.L.; D'Ambros,R.; Leite,R.C.; Heneimann,M.B.; Reis,J.K.P.
O objetivo deste trabalho foi desenvolver uma PCR em tempo real (qPCR) para o diagnóstico rápido e sensível da doença de Aujeszky. Os iniciadores amplificaram um fragmento de 123 pares de base do gene codificante da glicoproteína D. A qPCR foi testada em 25 amostras de cérebro de suíno positivas e 85 amostras negativas para DA no isolamento viral e na soroneutralização. A sensibilidade analítica foi calculada com acréscimo de um isolado brasileiro do SuHV-1 titulado em amostras de cérebro de suíno negativas na soroneutralização e na PCR. A técnica apresentou sensibilidade analítica de 10-0,5 TCID50/50µL. A qPCR foi capaz de distinguir reações inespecíficas devido a dímero de oligonucleotídeos iniciadores ou amplificações, além do alvo designado (evitando,...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Suínos; QPCR; Validação; Doença de Aujeszky.
Ano: 2013
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352013000300028

Evaluation of Reference Genes for Quantitative PCR in Four Tissues from Rabbits with Hypercholesterolaemia
Provedor de dados: 52
Autores: Zhang,Zhen; Wen,Bin; Xu,Yuan; Jiang,En-ze; Liu,Jia-yu; Zhu,Ke-li; Ning,Fang-yong; Du,Zhi-Heng; Bai,Xiu-Juan.
Abstract Rabbit with hypercholesterolaemia is an important model for studying cholesterol metabolism disease. This study aimed to evaluate the expression stability of nine reference genes for quantitative PCR (qPCR) analysis in adrenal gland, liver, spleen, and kidney tissue from rabbits with hypercholesterolaemia. In total, 30 male Harbin Large White (HLW) rabbits were fed a normal feed (n = 15) or a high cholesterol feed (n = 15) for 8 weeks to induce hypercholesterolaemia. Nine reference genes were verified by qPCR using cDNA extracted from rabbit tissue samples. For qPCR analysis, reference genes were evaluated using the RefFinder and GeNorm algorithms. Overall, seven rabbits with hypercholesterolaemia were identified based on body weight and total...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Evaluation; Reference genes; QPCR; Rabbits; Hypercholesterolaemia.
Ano: 2019
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132019000100428

Evaluation of reactive Epstein-Barr virus (EBV) in Iranian patient with different subtypes of multiple sclerosis (MS)
Provedor de dados: 55
Autores: Ramroodi,Nourollah; Niazi,Abbas Ali; Sanadgol,Nima; Ganjali,Zohre; Sarabandi,Vida.
OBJECTIVES: Epstein-Barr virus has been recently associated with the onset of multiple sclerosis, yet understanding how it elicits autoimmunity remains elusive. We investigated the relation between Epstein-Barr virus reactivation and disease development in different subtypes of multiple sclerosis. METHODS: In the present research, we have determined the Epstein-Barr virus-DNA load by quantitative real-time polymerase chain reaction and Epstein-Barr virus antibody levels by EIA technique in both multiple sclerosis patients (n = 78) and healthy controls (n = 123). RESULTS: Our results demonstrated increased titer of both anti-Epstein-Barr virus-IgG and IgM antibodies in patients (91.02% vs 82.11% in controls, p &lt; 0.001 and 14.1% vs 4.06% in controls, p...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: EBV; MS; Reactivation; QPCR; ELISA.
Ano: 2013
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702013000200007

qPCR detection of Mycobacterium leprae in biopsies and slit skin smear of different leprosy clinical forms
Provedor de dados: 55
Autores: Azevedo,Michelle de Campos Soriani; Ramuno,Natália Mortari; Fachin,Luciana Raquel Vincenzi; Tassa,Mônica; Rosa,Patrícia Sammarco; Belone,Andrea de Faria Fernandes; Diório,Suzana Madeira; Soares,Cleverson Teixeira; Garlet,Gustavo Pompermaier; Trombone,Ana Paula Favaro.
Abstract Leprosy, whose etiological agent is Mycobacterium leprae, is a chronic infectious disease that mainly affects the skin and peripheral nervous system. The diagnosis of leprosy is based on clinical evaluation, whereas histopathological analysis and bacilloscopy are complementary diagnostic tools. Quantitative PCR (qPCR), a current useful tool for diagnosis of infectious diseases, has been used to detect several pathogens including Mycobacterium leprae. The validation of this technique in a robust set of samples comprising the different clinical forms of leprosy is still necessary. Thus, in this study samples from 126 skin biopsies (collected from patients on all clinical forms and reactional states of leprosy) and 25 slit skin smear of leprosy...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Mycobacterium leprae; QPCR; Leprosy; Bacilloscopy.
Ano: 2017
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702017000100071

Development and validation of a modified TaqMan based real-time PCR assay targeting the lipl32 gene for detection of pathogenic Leptospira in canine urine samples
Provedor de dados: 58
Autores: Miotto,Bruno Alonso; Hora,Aline Santana da; Taniwaki,Sueli Akemi; Brandão,Paulo Eduardo; Heinemann,Marcos Bryan; Hagiwara,Mitika Kuribayashi.
Abstract A modified TaqMan real-time polymerase chain reaction targeting a 138 bp fragment within the lipl32 gene was developed to identify exclusively pathogenic Leptospira spp. in dog urine samples. Thirty-five samples from dogs with suspected clinical leptospirosis and 116 samples from apparently healthy dogs were tested for presence of leptospiral DNA using the TaqMan-based assay. The results were compared with those from a well-established conventional PCR targeting the 16S RNA encoding gene associated with nucleotide sequencing analysis. The overall agreement between the assays was 94.8% (confidence interval [CI] 95% 88-100%). The newly developed assay presented 91.6% (CI 95% 71.5-98.5%) relative sensitivity (22[+] lipl32 PCR/24[+] 16S RNA and...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Leptospirosis; Dog; Urine; QPCR; Lipl32.
Ano: 2018
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822018000300584

Seasonal variation on the presence of adenoviruses in stools from non-diarrheic patients
Provedor de dados: 58
Autores: Vetter,Michele Regina; Staggemeier,Rodrigo; Vecchia,Andréia Dalla; Henzel,Andréia; Rigotto,Caroline; Spilki,Fernando Rosado.
Human adenoviruses (HAdV), members of the Adenoviridae family, are excreted through the fecal route and may be present in the feces of humans consuming contaminated food or water. The presence of HAdV from different serotypes in the feces of healthy individuals was already reported using conventional polymerase chain reaction; however, real-time PCR (qPCR) may reveal not only the rates of detection as well as demonstrate the viral loads excreted by healthy persons. Aiming to identify and characterize the presence of adenoviruses in stool samples, 147 fecal samples from patients with no records of diarrhea were analyzed (74 from winter season and 73 from summer) by Real-Time PCR (qPCR) assay and conventional PCR. HAdV genome was present in 43.8% (32/73) of...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: HAdV; Stool samples; QPCR; Healthy individuals.
Ano: 2015
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822015000300749

Different methods to quantify Listeria monocytogenesbiofilms cells showed different profile in their viability
Provedor de dados: 58
Autores: Winkelströter,Lizziane Kretli; Martinis,Elaine C.P. De.
Listeria monocytogenes is a foodborne pathogen able to adhere and to form biofilms in several materials commonly present in food processing plants. The aim of this study was to evaluate the resistance of Listeria monocytogenes attached to abiotic surface, after treatment with sanitizers, by culture method, microscopy and Quantitative Real Time Polymerase Chain Reaction (qPCR). Biofilms of L. monocytogenes were obtained in stainless steel coupons immersed in Brain Heart Infusion Broth, under agitation at 37 °C for 24 h. The methods selected for this study were based on plate count, microscopic count with the aid of viability dyes (CTC-DAPI), and qPCR. Results of culture method showed that peroxyacetic acid was efficient to kill sessile L. monocytogenes...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Biofilms; L. monocytogenes; QPCR; Peroxyacetic acid; Sodium hypochlorite.
Ano: 2015
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822015000100231

In-house quantitative real-time PCR for the diagnosis of hepatitis B virus and hepatitis C virus infections
Provedor de dados: 58
Autores: Zauli,Danielle Alves Gomes; Menezes,Carla Lisandre Paula de; Oliveira,Cristiane Lommez de; Mateo,Elvis Cristian Cueva; Ferreira,Alessandro Clayton de Souza.
Abstract The quantification of viral nucleic acids in serum by real-time PCR plays an important role in diagnosing hepatitis B virus and hepatitis C virus infection. In this study, we developed an assay using specific primers and probes to quantify hepatitis B virus DNA or hepatitis C virus RNA in serum from infected patients. For standardization and validation of the assay, an international panel of hepatitis B virus/hepatitis C virus and standard plasmids was used. A correlation coefficient of 0.983 and 0.963 for hepatitis B virus and hepatitis C virus, respectively, was obtained based on cycle threshold values and concentrations of DNA or RNA. The standard curve showed a linear relationship from 19 IU/mL to 1.9 × 109 IU/mL of serum, with a coefficient...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: QPCR; Hepatitis B; Hepatitis C.
Ano: 2016
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822016000400987

Correlation between chronic inflammation, immunostaining and parasite load in the genital system of female dogs naturally infected with Leishmania infantum
Provedor de dados: 65
Autores: Oliveira,Vinícius Vasconcelos Gomes de; Macedo,Simone Regina Barros de; Araújo,Flábio Ribeiro de; Ramos,Carlos Alberto do Nascimento; Ramos,Rafael Antonio Nascimento; Maia,Frederico Celso Lyra; Alves,Leucio Câmara; Silva Junior,Valdemiro Amaro da.
ABSTRACT: The aim of the present study was to investigate the correlations among chronic inflammatory reaction, immunostaining and parasite load in the genital system of female dogs naturally infected with  Leishmania infantum . Animals (n = 10) used in this study were from the Department of Vector Control and Animal Surveillance of the municipality of Caruaru, state of Pernambuco, Brazil. Fragments of the vulva, vagina, cervix, uterine body, uterine horns and ovaries were submitted to histopathological analysis, immunohistochemistry (IHC) and DNA detection of amastigotes by qPCR. Correlations were found between the IHC findings and chronic inflammatory infiltrate related to L. Infantum only in the vulva and vagina; whereas, the same inflammatory reactions...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Canine visceral leishmaniasis; Genital pathology; Immunohistochemistry; QPCR.
Ano: 2016
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782016001102029

Associação da carga parasitária renal com achados laboratoriais em cães com leishmaniose visceral
Provedor de dados: 65
Autores: Torres,Mariana de Medeiros; Almeida,Arleana do Bom Parto Ferreira de; Sorte,Eveline da Cruz Boa; Paula,Daphine Ariadne Jesus de; Oliveira,Anderson Castro Soares de; Pescador,Caroline Argenta; Mendonça,Adriane Jorge; Nakazato,Luciano; Sousa,Valéria Régia Franco.
A leishmaniose visceral canina é uma doença grave e a morte ocorre por falência renal, considerando que os métodos diagnósticos convencionais não possibilitam a classificação clínica do animal. O objetivo deste estudo foi associar a carga parasitária renal aos achados clínicos e histopatológicos em cães com leishmaniose visceral. A análise microscópica revelou predomínio de nefrite intersticial mononuclear de graus variados em 59,3% dos cães avaliados. Entretanto, não houve diferença entre a carga parasitária renal de sintomáticos e oligossintomáticos (P= 0,35). As lesões renais foram de ordem inflamatória e a quantidade de parasitos não influenciaram na característica dessas lesões e nem nas alterações bioquímicas, mesmo em cães com diferentes...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Rim; QPCR; Cão; Leishmania sp.
Ano: 2013
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782013000500023

Feline herpesvirus 1 viral load related to environmental factors in sheltered cats
Provedor de dados: 65
Autores: Baumworcel,Natasha; Pereira,Joylson de Jesus; Soares,Ana Maria Barros; Souza,Guilherme Nunes; Almosny,Nadia Regina Pereira; Castro,Tatiana Xavier de.
ABSTRACT:  Shelter environment stress factors are related to FHV-1 viral reactivation. However, comparisons between conjunctival viral load and environmental factors have not been commonly evaluated. The aim of this study was to correlate FHV-1 viral load in domestic cats with and without clinical signs of conjunctivitis to shelter design in order to use FHV-1 viral load as a parameter of “health management”. Cats from four different shelters underwent an ophthalmological examination. Samples were collected by rolling a DNA/RNAse-free cytobrush over the ventral conjunctival fornix and were stored in 1.5 mL sterile microtubes in 500 μL of Eagle’s minimum essential medium and kept at 4 ºC. Molecular procedures were performed up to 48 hours after collection....
Tipo: Info:eu-repo/semantics/article
Palavras-chave: FHV-1; Conjunctivitis; Shelter medicine; QPCR; Animal shelter.
Ano: 2019
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782019000500454

PCR em tempo real para diagnóstico da leucose enzoótica bovina
Provedor de dados: 65
Autores: Dias,Natanael Lamas; Fonseca Júnior,Antônio Augusto; Rodrigues,Daniel Sobreira; Camargos,Marcelo Fernandes.
O objetivo deste trabalho foi realizar a validação de uma reação em cadeia da polimerase em tempo real com o sistema Plexor® (qPCR) para o diagnóstico da Leucose Enzoótica Bovina (LEB), por meio da comparação com testes de diagnóstico recomendados pela Organização Mundial de Saúde Animal (OIE). A qPCR foi comparada com duas outras técnicas: a PCR nested (nPCR) e a imunodifusão em gel de ágar (IDGA). Das 82 amostras analisadas pela qPCR e nPCR, 79 apresentaram resultados concordantes, sendo a concordância, classificada pelo Índice Kappa, como alta. Entre as PCRs e a IDGA, o número de resultados concordantes foi de 71 e 69, respectivamente, para qPCR e nPCR, sendo a concordância classificada como considerável. A qPCR apresentou altos valores de sensibilidade...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: BLV; Validação; QPCR; NPCR; IDGA.
Ano: 2012
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782012000800017

Expression of genes SBP and leginsulin in contrasting soybean seed coats
Provedor de dados: 65
Autores: Bahry,Carlos André; Zimmer,Paulo Dejalma.
ABSTRACT: Evaluation of differential candidate gene expression in contrasting soybean seeds is an auxiliary tool in the partial elucidation of processes involved in seeds formation, as well as it contributes to the generation of new information that can be used in future research or in the development of r genetic superior constitutions. The aim of this study was to evaluate the expression of two candidate genes, SBP and leginsulin genes, possibly involved in seed quality, in contrasting coats of four soybean genotypes. Two cultivars of yellow soybeans were used, BMX Potência RR and CD 202, and two lines of black soybean, TP and IAC. Gene expression was evaluated using qPCR in seven stages of development from seed coats for four genotypes, at 25, 30, 35,...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Gene expression; Soybean coats; QPCR.
Ano: 2016
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782016001001695